Extracellular Hsp70 (eHsp70) exerts its biological actions via Toll‐like receptors 2 and 4, and is increased in sera of chronic obstructive pulmonary disease (COPD) patients. The aim of this study was to explore the pro‐inflammatory effects and cytotoxicity of eHsp70 alone and in combination with bacterial components lipoteichoic acid (LTA) and lipopolysaccharide (LPS) on NCI‐H292 airway epithelial cells. NCI‐H292 cells were treated with recombinant human Hsp70 protein (rhHsp70), LPS, LTA and their combinations for 4, 12, 24 and 48 hours. IL‐6, IL‐8 and TNF‐α levels were measured by an ELISA method. Cell viability was determined by the MTS method, and caspase‐3/7, caspase‐8 and caspase‐9 assays. rhHsp70 induced secretion of IL‐6 and IL‐8 in a concentration‐ and time‐dependent manner, with the highest secretion at 24 hours. rhHsp70 combined with LTA had antagonistic and with LPS synergistic effect on IL‐6 secretion, while the interactions between rhHsp70 and LPS or LTA on IL‐8 were synergistic. TNF‐α was not detected in the applied conditions. rhHsp70, LPS or LTA did not affect cell viability, and rhHsp70 even suppressed caspase‐3/7 activities. We suggest that pro‐inflammatory effects of eHsp70, together with other damaging molecules and/or COPD risk factors, might contribute to the aggravation of chronic inflammation in human bronchial epithelium.

中文翻译：

---

细胞外Hsp70对NCI-H292人支气管上皮细胞系的促炎作用。

细胞外Hsp70（eHsp70）通过Toll样受体2和4发挥其生物学作用，并在慢性阻塞性肺疾病（COPD）患者的血清中增加。这项研究的目的是探讨单独使用eHsp70并与细菌成分脂磷壁酸（LTA）和脂多糖（LPS）联合使用对NCI-H292气道上皮细胞的促炎作用和细胞毒性。用重组人Hsp70蛋白（rhHsp70），LPS，LTA及其组合处理NCI-H292细胞4、12、24和48小时。IL-6，IL-8和TNF-α水平通过ELISA方法进行测量。细胞活力通过MTS方法以及caspase-3 / 7，caspase-8和caspase-9测定来确定。rhHsp70以浓度和时间依赖性方式诱导IL-6和IL-8的分泌，其中24小时的分泌最高。rhHsp70联合LTA对IL-6分泌具有拮抗作用和LPS协同作用，而rhHsp70与LPS或LTA对IL-8的相互作用具有协同作用。在应用条件下未检测到TNF-α。rhHsp70，LPS或LTA不会影响细胞活力，rhHsp70甚至抑制caspase-3 / 7活性。我们认为eHsp70的促炎作用以及其他破坏性分子和/或COPD危险因素可能会加剧人支气管上皮的慢性炎症。